The immune system protects an organism against diseases by identifying and eliminating the pathogen as well as suppressing the emergence of chronic diseases, such as tumors. In addition, the immune system participates in processes that maintain stable conditions (homeostasis) during development and growth, and following inflammatory reaction or tissue damage. Immune system is classically divided into the innate and the adaptive systems. Adaptive immune system of vertebrate species including human is supplied with an ability to memorize the fingerprints of hostile events threatening organism's physical integrity. The complex immune memory is built upon specialized cells and molecules seeking and destroying disease fingerprints that can be defined as antigens characteristic to either pathogens or structures interpreted as pathogenic (Paul 1993). During the antibody recognition, the immune cells produce antibodies serving as effector molecules against antigens.
Immunoglobulins (Ig) i.e. antibodies are defined as sensory molecules able to detect foreign insult or hostile invaders i.e pathogens and tag them for elimination, and in that way protecting organism against the disease. Next to hostile invaders, such as microbes (virus or bacteria) or parasites threatening the integrity of the host organism, a vaccine that mimics microbial invasion, a host's own naturally occurring molecular entity that has served up it's natural function, or an allergen are considered as immune insults (Berzofsky et al 1993). Also, under special circumstances, the normal constituents of the body can be targeted by the immune system hence can be considered a hostile structures (Suber et al 2008). The fractions of Ig represented in mammals constitute of IgG, IgM, IgE, IgA, and IgD, synthesized by B-lymphocytes in response to antigenic stimuli.
Immunoglobulins are composed of two light and heavy chains that form a Y-like structure that binds antigens with the end of their arms, a structure called paratope. Relevant to the current invention, the antigens recognized by a paratope can be determined by reverse engineering. Molecular libraries that mimic the structure of antigen epitopes including proteins, oligosaccharides, lipopolysaccharides (LPS), glycosphingolipids (gangliosides), and other types of organic and inorganic molecules can be used to characterize naturally occurring epitopes targeted by Igs (Berzofsky 1993). For characterization, various technologies including ELISA, RIA, and interaction display methods such as phage display can be used. Peptide substitution analysis and random display mapping experiments have shown that core B-cell epitopes are relatively short ranging between 3-12 amino acids, borders of the mapped epitope are sharp and only some residues are critically required for interaction (Buus et al 2012). Different antibody epitope mapping methods have reported, yet as a rule, the single amino acid resolution epitope profiles for most of the monoclonal and polyclonal antibodies are not commonly available. There is a need for simple, robust, high-throughput, accurate and cost-effective epitope mapping method, enabling to map linear and conformational epitopes of mono- and polyclonal antibodies.
Igs play an important role in both health and disease, and, thus, their occurrence in the body reveals relevant clues about the health condition. The invention is based on the following statements: (1) Ig composition undergoes disease-specific changes, (2) the changes and shifts are preserved in the immune memory, and (3) the recorded events, if interpreted in the right way, can be used to make decisions about the health of the ogranism.
It has been demonstrated, that antibodies as well as antigens are useful in clinical immunodiagnostics, differential diagnostics, determination of prognosis, and informed decision-making about treatment options. To this end there also remains a need to develop methods to further characterize the immune system.
Traditional clinical immunodiagnostics is based on blood tests that apply circulating Igs as the detectors of antigens, or the precise copies of previously known antigens as detectors of seroreactivity. However obtaining either antibodies with specific affinity or whole antigenic molecules is costly and labor-intensive.
The invention describes a method that by generating a metadatabase enables to detect any immune memory-related health condition of an organism by catalogizing information related to the organism, related but not limited to the patterns of macromolecules that are structurally similar to naturally occurring disease antigens. With the method huge amounts of data of Ig minimal epitopes reflecting antigenic regions of the known and yet to be defined, novel substances are defined and simultaneously provided compiling a metadatabase. As the presence and levels of these minimal epitope-defining sequences can be measured all at once in a massively paralleled manner, eg numerous samples simultaneously (in DESCRIPTION OF INVENTION), the qualitative and quantitative variation of the minimal epitope defining sequences identified by the method, generate characteristic to an individual sample profiles that ultimately contribute to defining the antibody-activity related antigenic signatures reflecting the health medical conditions. If a statistically significant match with a disease or disorder is found, a diagnosis or other medical decision can be proposed. The invention is termed as Mimotope Variance Analysis (MVA).